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진단검사의학과곽호석

Tobacco smoke

• Tobacco smoke component: > 5,000 chemi-

cals

• Electronic database of smoke components:

>2,000 chemicals

–Mainstream smoke: 542 chemicals

• Human inhalation risk: 98 chemi-

calsA list of smoke components needs to be selected with a suffi-ciently broad chemical, toxicological, and pharmacological pro-file.

List of harzardous tobacco smoke compo-nents

Int. J. Environ. Res. Public Health 2011, 8

Int. J. Environ. Res. Public Health 2011, 8

Hazardous tobacco smoke component

• The list of Hoffmann analytes: biolog-

ically or toxicologically active main-

stream smoke components

– Based on research from the early 1990s

– Only includes carcinogens

The list of Hoffmann analytesGroup Analytes Average Yields

Aromatic Amines

1-Aminonaphthalene (ng/cig) 15.1

2-Aminonaphthalene (ng/cig) 10.3

3-Aminobiphenyl (ng/cig) 3

4-Aminobiphenyl (ng/cig) 1.7

Carbonyls

Methyl ethyl ketone (μg/cig) 62.72

Acetaldehyde (μg/cig) 560.48

Acetone (μg/cig) 264.74

Acrolein (μg/cig) 58.77

Butyraldehyde (μg/cig) 29.58

Crotonaldehyde (μg/cig) 16.18

Formaldehyde (μg/cig) 21.61

Propionaldehyde (μg/cig) 43.92

Phenols

Catechol (μg/cig) 37.9

Hydroquinone (μg/cig) 32.4

m+p-Cresol (μg/cig) 5.84

o-Cresol (μg/cig) 1.89

Phenol (μg/cig) 7.32

Resorcinol (μg/cig) 0.91

Polycyclic Aromatic Hydrocarbon (PAH)

Benzo[a]pyrene (ng/cig) 7

Group Analytes Average Yields

Inorganics

Ammonia (μg/cig) 11.02

Hydrogen Cyanide (μg/cig) 109.2

Nitric Oxide (μg/cig) 223.41

Carbon Monoxide (mg/cig) 11.96

Organics Acrylonitrile (μg/cig) 8.28

Volatile Hydrocarbons

1,3-Butadiene (μg/cig) 29.94

Benzene (μg/cig) 43.39

Isoprene (μg/cig) 297.68

Toluene (μg/cig) 64.91

Styrene (μg/cig) 5.11

Nitrogen Hetero-cyclics

Pyridine (μg/cig) 7.02

Quinoline (μg/cig) 0.23

Nicotine (mg/cig) 0.75 (750ug)

Metals & Metalloids

Arsenic (ng/cig) 10.4

Cadmium (ng/cig) 47.8

Chromium (ng/cig) 73

Lead (ng/cig) 33

Mercury (ng/cig) 3.82

Nickel (ng/cig) 5.12

Selenium (ng/cig) 34.9

Tobacco Specific Nitrosamines (T-

SNAs)

NAB (ng/cig) 16.3 (0.016ug)

NAT (ng/cig) 119 (0.119ug)

NNK (ng/cig) 115.6 (0.116ug)

NNN (ng/cig) 133.1 (0.133ug)

'Tar' Nicotine-free-dry-particu-late-matter (mg/cig) 8.91

Biomarkers of tobacco smoke ex-posure

• Carbon monoxide (CO)• Nicotine metabolite: Cotinine• Polycyclic aromatic hydrocarbon

(PAHs) metabolite: 2-naphthol, 3-hy-droxyfluorenes, 1-hydroxypyrene

• Tobacco specific nitrosamines (TSNAs) metabolite: NNAL

Tobacco-specific nitrosamines(TSNAs)

Processed tobaccos are invariably contaminated with TSNAs.

Formed during processing (drying and fermentation)

Not present in fresh and green tobaccos

Different types of tobacco contain different concentrations of ni-

trosamines

The higher the nitrate content of tobaccos the higher also the nitrosamine con-

tamination

The high-nitrate tobaccos or other dark tobacco types are high in TSNAs

No correlation between the declared tar content of cigarettes and

the TSNA concentration in mainstream tobacco smoke

No evidence that nicotine is nitrosated during the smoking process

Range for tar, nicotine and tobacco specific ni-trosamine (TSNA) in tobacco for commercial ciga-

rettes from several countries

Results of testing for NNK yields from three brands of cigarettes in various countries

We see these results as a compelling and urgent argument for government regulation of carcinogen concentrations in ciga-rettes.

Tobacco Control 2000;9:351–354

Nicotine yield range

0.85 mg/cig to 1.3 mg/cig 0.68 mg/cig to 1.25 mg/cig

Chemical structure of TSNAs

Nitrosation

Nitrosamine structure

Formation of tobacco-specific nitrosamines in tobacco

Potential third-hand smoke

PNAS ∣ April 13, 2010 ∣ vol. 107 ∣ no. 15

The metabolic activation and inactivation of NNK

* 11β-HSD: 11β-hydroxysteroid dehydrogenase

Steroid hormonesDrug: diureticsDrug: block 11β-HSDDrugDrug: flavonoidFlavanone

Similar structure: hormones

Chemical structure of NNAL

11β-HSD

Metabolism of NNK and NNAL and formation of adducts

Structures of characterizedpyridyloxobutyl DNA adducts

Proposed activation pathways for NNN, NNK and NNKOAc

Postulated metabolic pathway medi-ating the toxicity of NNK

Other chemi-cal in smoke

Concentrations of total urinary 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL) (pg/mg creatinine) and ciga-

rettesper day in 1054 National Health and Nutrition Examination

Survey2007–2008 smokers

Adjusted geometric means in regression model with 95%

confidence intervals.

The metabolic activation and inactivation of NNK

* 11β-HSD: 11β-hydroxysteroid dehydrogenase

Steroid hormonesDrug: diureticsDrug: block 11β-HSDDrugDrug: flavonoidFlavanone

Similar structure: hormones

Relationships between Cigarette Con-sumption and Biomarkers of Tobacco

Toxin ExposureCO

NNAL Cotinine

1-HOP (PAHs metabolite)

Structures of NNK, NNAL, and NNAL-Gluc

Quantitation of Metabolites of NNK af-ter Cessation of Smokeless Tobacco Use

Time post-cessation (days)

Smokeless tobacco Smoking

NNAL t1/2: 10-18 days > cotinine t1/2 : 1days

Metabolizer phenotypes for smok-ers

• PM: poor metabolizer (ratio<2)• IM: intermediate metabolizer (ratio between 2 and 5)• EM: extensive metabolizer (ratio >5)

Comparison of free, conjugated and total urinary NNAL for 60 smokers

NNAL as a Biomarker of exposure to cigarette smoke at the preg-

nancy

• Smoking patterns during pregnancy

• Second hand smoke during preg-

nancy

• Fetal exposure

Smoking patterns during preg-nancy

Exposure to different sources of second-hand smoke during pregnancy and its effect on urinary cotinine and NNAL con-

centrations

TOBACCO SMOKING OR EXPOSURE TO SECOND-HAND SMOKE ON THE LEVELS

OF NNAL IN URINE OF MOTHER AND NEWBORN

Development of LC-MS/MS for NNAL in urine

• Simple sample preparation

• Rapid analytical time

• High sensitivity

Enzyme treat• NNAL-glucuronides hydrol-ysis

• Time: 20h

Extraction• Liquid-Liquid extraction• Time: 1h

Derivatization• N-propyl derivatization• Time: 12h

Analysis LC-MS/MS• Analytical time: 30min / test

- Sample preparation

LC-MS/MS analysis of NNAL

Blank Urine

LLOQ: 20 pg/mL

LC-MS/MS analysis of NNALBefore enzyme treatment

After enzyme treatment

NNAL-glucuronides Concentration: NNAL concentration (After enzyme treatment) – NNAL (Before enzyme treatment)

• Simple sample preparation

– Each detection of NNAL and NNAL-glucuronides

• Rapid analytical time

– Optimization of analytical column

• High sensitivity

– Optimization of sample preparation

The End